Methanol fixed fibroblasts serve as feeder cells to maintain stem cells in the pluripotent state in vitro.

ABSTRACT: Preparation of mouse embryonic fibroblast (MEF) feeder cells to maintain pluripotent stem cells (PSCs) is time consuming and involved in animal issues. Here, we demonstrated a novel method to prepare feeder cells with high efficiency, timesaving, and low costs. MEFs in 3 × 10⁴ cell/cm² were fixed by methanol for 5 min and air drying for 5 min. Thereafter, the methanol fixed MEF cells (MT-MEF) were able to be used directly to culture PSCs or stored at room temperature for the future usage. PSCs cultured on MT-MEF could be continuously expanded for over 40 passages with the naïve pluripotency. MT-MEFs could also be used to maintain human and pig iPSCs. Moreover, methanol fixed MEFs' culture dish was able to be reused for at least 4 times, and to be applied for antibioti

SUBMITTER: Ren Y 

PROVIDER: S-EPMC5958091 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature